TR&D2: Biomedical Research Utilizing Dynamic Nuclear Polarization Project Summary/Abstract The development of dynamic nuclear polarization (DNP) to increase the sensitivity of NMR by 1-2 orders of magnitude is profoundly impacting the field of NMR. Early results demonstrating the utility of DNP when combined with magic angle spinning (MAS) solid state NMR (ssNMR) for biomolecular studies provide impetus for further developing DNP technology to support biomedical research applications. This TR&D addresses challenges the NMR community faces in making DNP a truly orthogonal, enabling technology for biomolecular ssNMR through three specific aims: 1) develop DNP MAS ssNMR probes with flexible electronic architecture, fast MAS, and extended VT Range; 2) develop a biradical toolbox for targeted DNP enhancement of specific protein classes; and 3) optimize DNP MAS ssNMR approaches for biomolecular structure determination. The goal of these aims is to provide a holistic approach to maximizing the promise of DNP through hardware development, synthesis of needed chemical reagents, optimization of sample preparation strategies, and developing the experimental framework for biomolecular ssNMR in the age of DNP. Advances in all these areas must be pursued to gain the needed sensitivity and resolution for leading edge biomedical research applications and to transition DNP MAS ssNMR to a technique that is more widely accessible to the structural biology community as a whole. Through partnering with six Driving Biomedical Research projects, we will rapidly develop and deploy technology in this TR&D to address biomedical research problems in the areas of membrane protein structure determination, amyloid biogenesis in bacterial biofilms and in neurodenerative diseases, protein assembly in viruses, and enzyme catalysis.